This kit is optimized to detect West Nile Virus protease activity. It provides enough reagents to perform 100 assays in a 96-well format.
High Speed: The entire process only takes about half an hour.
West Nile virus (WNV) is from the family Flaviviridae3 found in both tropical andtemperate regions. The main route of human infection is through the bite of an infected mosquito Infection causes severe neurological disease and fatalities in both human and animal hosts. There is currently no effective vaccine or antiviral drug to protect against WNV infection.4WNV contains a single-stranded, positive-sense RNA genome, which encodes threestructural proteins [capsid (C), membrane (M), envelope (E)], and seven non-structural proteins(NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5).5,6 West Nile viral NS3 protease is absolutelyessential (along with viral-encoded cofactor NS2B) for post-translational processing of a viralpolypeptide precursor in infected host cells. This polypeptide provides the structural and functional viral proteins, and inhibition of its processing could represent a potential treatment for viral infections. Thus, this protease has become one of the key targets for developing anti-WNV drugs.1,7
The 570 West Nile Virus Protease Assay Kit provides a convenient,homogeneous assay for high throughput screening of West Nile Virus protease NS3 inhibitors. Itallows for continuous quantification of protease activity using QXL™570/5-TAMRA FRETpeptide. In the FRET peptide, the fluorescence of 5-TAMRA is quenched by QXL™570. Uponcleavage into two separate fragments by WNV protease, the fluorescence of 5-TAMRA isrecovered, and can be monitored at excitation/emission = 540 nm/575 nm. The long wavelength fluorescence of 5-TAMRA is less interfered by the autofluorescence of cell components and test compounds.
Materials Required (but not provided): • WNV protease: WNV protease can be produced in an E. coli expression system provides active recombinant WNV protease
96-Well microplate: Black microplates can provide better signal to noise ratio.
Fluorescence microplate reader: Capable of detecting emission at 575 nm with excitation at 540nm.
Storage and Handling:
Store all kit components at -20°C.
Protect Components A and B from light and from moisture.
Component C can be stored at room temperature for convenience.
